Global Protein Profiling in Processed Immunohistochemistry Tissue Sections.
Simone VenzViola von Bohlen Und HalbachChristian HentschkerHeike JunkerAndreas Walter KussThomas SuraElke KrügerLinus VölkerOliver von Bohlen Und HalbachLars Riff JensenElke HammerPublished in: International journal of molecular sciences (2023)
Tissue sections, which are widely used in research and diagnostic laboratories and have already been examined by immunohistochemistry (IHC), may subsequently provide a resource for proteomic studies, even though only small amount of protein is available. Therefore, we established a workflow for tandem mass spectrometry-based protein profiling of IHC specimens and characterized defined brain area sections. We investigated the CA1 region of the hippocampus dissected from brain slices of adult C57BL/6J mice. The workflow contains detailed information on sample preparation from brain slices, including removal of antibodies and cover matrices, dissection of region(s) of interest, protein extraction and digestion, mass spectrometry measurement, and data analysis. The Gene Ontology (GO) knowledge base was used for further annotation. Literature searches and Gene Ontology annotation of the detected proteins verify the applicability of this method for global protein profiling using formalin-fixed and embedded material and previously used IHC slides.
Keyphrases
- mass spectrometry
- tandem mass spectrometry
- protein protein
- data analysis
- white matter
- amino acid
- liquid chromatography
- resting state
- binding protein
- high performance liquid chromatography
- systematic review
- single cell
- high resolution
- type diabetes
- genome wide
- gas chromatography
- functional connectivity
- small molecule
- simultaneous determination
- adipose tissue
- ultra high performance liquid chromatography
- electronic health record
- multiple sclerosis
- ms ms
- cognitive impairment
- atomic force microscopy
- solid phase extraction